CN111996231A - Method for identifying wheat scab expansion resistance by inoculating double flowers at top of wheat head - Google Patents
Method for identifying wheat scab expansion resistance by inoculating double flowers at top of wheat head Download PDFInfo
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- CN111996231A CN111996231A CN202010840850.8A CN202010840850A CN111996231A CN 111996231 A CN111996231 A CN 111996231A CN 202010840850 A CN202010840850 A CN 202010840850A CN 111996231 A CN111996231 A CN 111996231A
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Abstract
The invention discloses a method for identifying wheat scab extension resistance by double-flower inoculation at the top of a wheat ear. The disease part of the inoculation method is mainly expanded downwards from the sixth spikelet at the top, the disease spikelet rate at the lower part of the sixth spikelet is counted, the disease parts are consistent, the problem that identification reliability is affected due to withered spikelets easily caused by a traditional single flower drip method is solved, and the expansion resistance of the variety can be reflected more accurately.
Description
Technical Field
The invention relates to a method for identifying wheat scab expansion resistance by inoculating double flowers at the top of a wheat ear, belonging to the technical field of plant disease resistance identification and evaluation.
Background
The method for identifying the gibberellic disease resistance extension at present is mainly single-flower instillation method identification, namely, single florets of middle and lower spikelets of wheat are injected with spore liquid in the flowering period of wheat, the inoculation time is recorded, and the disease spikelet rate is counted 18-21 days after inoculation, namely, the disease resistance evaluation is carried out on the disease spikelet percentage in the total spikelet number. The traditional single flower drip method for inoculating the spikelet has the middle and lower parts of the spike, the phenomenon of withered spikelet is often caused by the fact that the upper part of the spikelet is lack of water and nutrients due to the fact that the vascular bundle is damaged, and particularly in genetic research, the withered spikelet is the main reason for causing large errors in gibberellic disease identification and difficulty in accurately positioning genes.
Disclosure of Invention
The invention aims to solve the problems of easy occurrence of withered and white ears and poor consistency of identification results in the traditional single flower drip method, and provides the gibberellic disease expansion resistance identification method which is clear in inoculated spikelet position, high in inoculation success rate, obvious in resistance phenotype difference and good in disease consistency.
In order to solve the technical problem, the invention provides a method for identifying the wheat scab expansion resistance by double-flower inoculation at the top of a wheat ear.
Further, the specific inoculation method comprises the following steps: inoculating the prepared gibberellin spore liquid between the inner glumes and the outer glumes of the florets on the two sides of the sixth spikelet on the top of the wheat ear by using a medical injector and an injection needle, attaching a waterproof label to the position, close to the base spikelet, below the ear, and recording the inoculation date and the strain name on the label.
Further, the inoculation amount of the gibberella spore liquid is 20 uL.
Further, the preparation method of the gibberella spore solution comprises the following steps: use of discs from activated solid plate Medium of strains of gibberellinTaking 3-5 PDA solid culture medium with gibberellin strain, putting into 60mL sterilized CMC culture medium, then putting on a shaker, shaking at 180rpm and 25-28 deg.C for 72-120 hr, sampling, observing spore production condition, sucking 10uL spore liquid, dripping onto a blood count plate, measuring spore concentration under microscope, and diluting or concentrating to 105spores/mL.
Furthermore, the recorded browning condition of the inoculated spikelets refers to that the browning condition of the inoculated spikelets is observed and recorded 3-5 days after inoculation, and the spikelets which are not successfully inoculated are removed.
Furthermore, the disease spikelet rate of the lower susceptible spikelet of the inoculated spikelet is counted respectively on the 14 th day and the 21 th day from the next day of inoculation, in order to eliminate the phenomenon of withered white spike which often occurs in the middle and lower part inoculation, the identification error is reduced, and the accuracy of the identification of the wheat scab expansion resistance is improved.
Furthermore, the ear-sick rate refers to the ratio of the number of susceptible ears to the total number of ears.
The main technical indexes for judging the quality of the gibberellic disease expansion resistance evaluation method system comprise inoculation success rate and stability of phenotype identification;
compared with the difference between the traditional single-flower drip method for inoculating the wheat head to the middle part and the lower part of the wheat head and the 2 technical indexes, the method has the advantages and positive effects that:
(1) according to the traditional single-flower drip method, spore liquid is injected into a single floret in the middle ear and the lower spikelet, and the inoculation part is not fixed;
(2) the invention adopts double-flower drip infusion, so the inoculation success rate is higher compared with the traditional single-flower drip infusion method;
(3) the disease part of the inoculation method of the invention mainly expands downwards from the sixth spikelet at the top, and counts the lower diseased spikelet rate of the sixth spikelet, the disease parts are consistent, and the problem that the traditional single-flower drip method easily causes spikelet withering and affects the identification reliability is solved, and the expansion resistance of the variety can be more accurately reflected.
Detailed Description
The invention is further described below. The following examples are only for illustrating the technical solutions of the present invention more clearly, and the protection scope of the present invention is not limited thereby.
The invention is used for carrying out the dynamic identification of gibberellic disease inoculation and expansion resistance on a gibberellic disease resistant strain R75 and a gibberellic disease susceptible strain S98 in the field. And (4) dynamically counting the sick spikelet rate of the lower susceptible spikelet of the inoculated spikelet according to the 14 th day and the 21 st day from the next day of inoculation, thereby identifying the expansion resistance of the variety.
The method for identifying the wheat scab expansion resistance comprises the following steps: using a disc sampler from an activated solid plate culture medium of the strain of the gibberellin to take 3 to 5 blocks of PDA solid culture medium with the strain of the gibberellin to be put into 60mL of sterilized mung bean soup culture medium, then putting the culture medium on a shaking table, shaking the culture medium for 72 to 120 hours at the temperature of 25 to 28 ℃ at 180rpm, sampling, observing the sporulation condition, sucking 10uL of spore liquid, dripping the spore liquid on a blood counting plate, measuring the spore concentration under a microscope, and then diluting or concentrating the spore liquid to the required concentration of about 105spores/mL. And (3) inoculating the prepared gibberella spore liquid between the inner glumes and the outer glumes of the small flowers on the two sides of the sixth small ear at the top of the wheat ear by using a medical injector and an injection needle in the flowering period of the wheat, attaching a waterproof label to the position close to the small ear at the base part between the lower sections of the wheat ear, and recording the inoculation date and the strain name on the label. Browning of the inoculated spikelets was observed and documented 3-5 days after inoculation. And (5) dynamically counting the sick spikelet rate of the lower susceptible spikelet of the inoculated spikelet at 14 th and 21 th days after inoculation, and identifying the wheat scab expansion resistance.
The method is used in fields for performing gibberellic disease inoculation and dynamic identification of expansion resistance, the disease spikelet rate of lower susceptible spikelets of inoculated spikelets is dynamically counted according to days 14 and 21 after inoculation to identify the variety expansion resistance difference, and experimental verification shows that the obtained result is that:
(1) inoculating and browning time of spikelets: the browning condition of inoculated spikelets is an important index for judging whether inoculation is successful or not, browning occurs if inoculation is successful, browning does not occur if inoculation is failed, and in the latter case, the spikelet rate is not counted dynamically, and R75 and S98 both appear in the browning condition at the inoculation point 3-5 days after inoculation, and no difference exists among strains.
(2) Lesion spikelet rate at day 14 post inoculation: the difference in the resistance to expansion between varieties gradually increases. The spikelet rates of single spike disease inoculated by R75 are 0.05, 0.16 and 0.16, and the average spikelet rate of disease is 0.12; and the ear disease rate of S98 is 0.32, 0.27 and 0.27 respectively, and the average ear disease rate is 0.29.
(3) Ear rate of disease at day 21 post inoculation: the spikelet rates of single spike disease inoculated by R75 are 0.11, 0.21 and 0.32, and the average spikelet rate of disease is 0.21; and the ear disease rate of S98 was 0.77, and 0.77, respectively, and the average ear disease rate was 0.77. At this time, the difference of the expansion resistance among varieties is obvious, and the phenomenon of withered and white ears does not occur, so that the scab resistant phenotype can be determined.
The experimental results prove that the method for identifying the wheat scab expansion resistance has the advantages of definite inoculation spike, high inoculation success rate, high consistency of diseased parts, capability of avoiding the withered white spike phenomenon which is easy to occur in the traditional single flower drip method, stable and high accuracy of the identification result and capability of more accurately reflecting the expansion resistance of the variety.
The above description is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, several modifications and variations can be made without departing from the technical principle of the present invention, and these modifications and variations should also be regarded as the protection scope of the present invention.
Claims (7)
1. A method for identifying wheat scab expansion resistance by double-flower inoculation at the top of a wheat ear is characterized in that a double-flower instillation method is utilized at a wheat flowering stage, a prepared gibberellic disease spore solution is inoculated to bilateral florets of a sixth spikelet at the top of the wheat ear, the browning condition of the inoculated spikelet is recorded, the diseased spikelet rate of infected spikelets at the lower part of the inoculated spikelet is counted after inoculation, and the wheat scab expansion resistance is dynamically evaluated.
2. The method for identifying the wheat scab expansion resistance by the double-flower inoculation at the top of the wheat ear as claimed in claim 1, wherein the specific inoculation method is as follows: inoculating the prepared gibberellin spore liquid between the inner glumes and the outer glumes of the florets on the two sides of the sixth spikelet on the top of the wheat ear by using a medical injector and an injection needle, attaching a waterproof label to the position, close to the base spikelet, below the ear, and recording the inoculation date and the strain name on the label.
3. The method for identifying the wheat scab expansion resistance by double-flower inoculation on the top of wheat head as claimed in claim 1, wherein the inoculation amount of the gibberellic spore solution is 20 uL.
4. The method for identifying the wheat scab expansion resistance by the double-flower inoculation at the top of the wheat ear as claimed in claim 1, wherein the preparation method of the gibberellic spore solution is as follows: using a disc sampler from an activated solid plate culture medium of the strain of the gibberellin to take 3 to 5 blocks of PDA solid culture medium with the strain of the gibberellin to be put into 60mL of sterilized CMC culture medium, then putting the CMC culture medium on a shaker, shaking the CMC culture medium for 72 to 120 hours at the temperature of 25 to 28 ℃ under the condition of 180rpm, sampling, observing the sporulation condition, sucking 10uL of spore liquid, dripping the spore liquid on a blood counting plate, measuring the spore concentration under a microscope, and then diluting or concentrating the spore liquid to 105spores/mL.
5. The method for identifying the wheat scab spread resistance by double-blossom inoculation on the top of wheat ear as claimed in claim 1, wherein said recording the browning of the inoculated ear is observed 3-5 days after inoculation, and the ear which is not inoculated successfully is removed.
6. The method for identifying the wheat scab spread resistance by double-blossom inoculation on the top of wheat head as claimed in claim 1, wherein the diseased spikelet rate of the lower susceptible spikelet of the inoculated spikelet is counted respectively at 14 days and 21 days from the next day of inoculation.
7. The method for identifying the wheat scab spread resistance by double blossom inoculation on top of wheat head as claimed in claim 6, wherein said disease spikelet rate is the ratio of susceptible spikelets to total spikelets.
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| CN202010840850.8A CN111996231A (en) | 2020-08-20 | 2020-08-20 | Method for identifying wheat scab expansion resistance by inoculating double flowers at top of wheat head |
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| CN202010840850.8A CN111996231A (en) | 2020-08-20 | 2020-08-20 | Method for identifying wheat scab expansion resistance by inoculating double flowers at top of wheat head |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113424720A (en) * | 2021-06-29 | 2021-09-24 | 西藏自治区农牧科学院农业研究所 | Identification method for loose smut resistance of highland barley |
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| CN104303866A (en) * | 2014-11-18 | 2015-01-28 | 扬州大学 | Method for identifying and evaluating wheat scab expansion resistance |
| CN105850537A (en) * | 2016-04-06 | 2016-08-17 | 江苏省农业科学院 | Identification and evaluation method for wheat scab extension resistance |
| CN111020000A (en) * | 2019-12-10 | 2020-04-17 | 扬州大学 | Method for identifying wheat scab resistance |
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- 2020-08-20 CN CN202010840850.8A patent/CN111996231A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104303866A (en) * | 2014-11-18 | 2015-01-28 | 扬州大学 | Method for identifying and evaluating wheat scab expansion resistance |
| CN105850537A (en) * | 2016-04-06 | 2016-08-17 | 江苏省农业科学院 | Identification and evaluation method for wheat scab extension resistance |
| CN111020000A (en) * | 2019-12-10 | 2020-04-17 | 扬州大学 | Method for identifying wheat scab resistance |
Non-Patent Citations (2)
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| 张语卉: ""小麦抗赤霉病主效QTL Fhb1在活体和离体条件下的效应解析"", 《中国优秀硕士学位论文全文数据库（电子期刊）农业科技辑》 * |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113424720A (en) * | 2021-06-29 | 2021-09-24 | 西藏自治区农牧科学院农业研究所 | Identification method for loose smut resistance of highland barley |
| CN113424720B (en) * | 2021-06-29 | 2023-06-09 | 西藏自治区农牧科学院农业研究所 | Identification method of highland barley anti-loose smut |
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